Abstract Foodborne illnesses devastate the gastrointestinal microflora of millions of Americans each year, often resulting in dysbiosis and increase risk of other diseases, namely irritable bowel syndrome, kidney failure, pancreatitis and diabetes. Furthermore, diabetics are at increased risk of developing a foodborne illness, and they often take longer to recover. Each year, 1 in 6 people in the U.S. are sickened by foodborne pathogens. In 2014, there were 19,542 laboratory confirmed infections, 4,445 hospitalizations and 71 deaths attributed to the 9 major pathogens transmitted commonly through food. Listeria is the one of the most serious offenders, with a 96% hospitalization rate and a 12.9% mortality rate. The impact on healthcare, business and industry is no less pronounced, resulting in annual costs of $14.1 to $16.3 billion, including direct medical costs and value of time lost to illness. Moreover, the cost of keeping food safe from Listeria contamination was estimated to range between $2.4-$2.6 billion. Contaminated food is the cause of these illnesses, having entered the public supply due to a lack of detection at a manufacturing/packaging plant, distribution warehouse or retail location. Current tests are inadequate because they require a prolonged enrichment step to reduce the likelihood of false negatives and false positives. This enrichment causes tremendous delays in providing actionable results to the food processor, who are left with one of two choices: Store the products until the test result comes back, resulting in older, lower-quality food; or shipping the food before receiving test results, putting the consuming public at increased risk. Sample6 is developing the world?s first enrichment-free foodborne pathogen detection system. DETECT/L is a rapid screening assay that detects a single Listeria spp. cell on environmental surfaces in less than one work shift (<7h). Sample6?s proprietary Bioillumination Platform enables this expeditious turnaround, enabling food manufacturers and packagers to quickly and accurately detect and remediate contaminated environmental surfaces, and solely retain product that is at risk of contamination. However, food can become contaminated internally during production, shipping, or in wholesale and retail environments downstream in the supply chain. It is of vital importance to develop a second testing modality that assays the foodstuff as rapidly and sensitively as DETECT/L does for environmental surfaces. To this end, the proposed Phase II study will continue the work begun in Phase I to adapt the DETECT/L assay for finished product testing. First, the researchers will expand the number of Listeria strains to 10. Next, assay development will be completed for the six foods begun in Phase I, plus 30 more foods. Finally, Sample6 will develop high-throughput methods so that food manufacturers and packagers can test multiple food samples at a time for efficiency.